Probiotic for infantile colic
AssigneesAB-Biotics, S.A., Venpharma Laboratorios, S. A.
InventorsJordi CUNE CASTELLANA, Elisabet LAZARO MALLEN, Jordi ESPADALER MAZO
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Abstract
The invention provides a bacterial composition which comprises from 104 to 1012 cfu/g of Pediococcus pentosaceus cells which have the ability to induce the production of interleukin-10 to reduce inflammation in the intestinal tract among other features. Thus, the bacterial composition is useful in the amelioration of excessive crying in infants. Particularly, Pediococcus pentosaceus cells are from the strain deposited as CECT 8330. The bacterial composition can be in the form of a food supplement, a medicament, an infant formula, an edible product and a food product. Particularly, the composition is in the form of an infant food supplement in the form of oily suspension.
Claims15
A freeze-dried bacterial composition which comprises from 104 to 1012 cfu/g of Pediococcus pentosaceus viable cells which induce the production of interleukin-10, wherein the production of interleukin-10 by THP-1 macrophages in the presence of Pediococcus pentosaceus cells as expressed as normalized increase is higher than the production of interleukin-10 by the negative control, which are THP-1 macrophages in the absence of Pediococcus pentosaceus cells, when the normalized increase is determined by the following steps:
- (a) differentiating THP-1 monocytes into macrophages by growing the THP-1 monocyte cell line obtained from the cell collection of the Public Health England, catalogue number 88081201, in Roswell Park Memorial Institute (RPMI) 1640 medium with 10% Fetal Bovine Serum (FBS), and with phorbol 12-myristate 13-acetate (PMA) to a final concentration of 0.16 μM;
- (b) growing the THP-1 macrophages in RPMI 1640 medium with 10% FBS in 24-wells ELISA plates to a final concentration of 106 macrophages/well;
- (c) incubating for 2.5 hours the THP-1 macrophages with lipopolysaccharide (LPS) at a final concentration of 10 ng/ml, and washing the THP-1 macrophages with Dulbecco's Phosphate Buffered Saline medium (D-PBS);
- (d) getting a culture of Pediococcus pentosaceus cells ready by having grown it overnight in Man, Rogosa and Sharpe medium (MRS) at 37° C. in a 5% CO2 atmosphere;
- (e) adding to each ELISA-well 500 μl of RPMI 1640 medium with 10% FBS and an appropriate amount of a dilution of Pediococcus pentosaceus cells to obtain a final ratio of 25:1, i.e. 2.5×107 cfu of Pediococcus pentosaceus cells:
- 106 THP-1 macrophages;
- (f) incubating the THP-1 macrophages with the Pediococcus pentosaceus cells for 2.5 hours at 37° C. or without the Pediococcus pentosaceus cells in the same conditions as negative control;
- (g) washing the THP-1 macrophages with D-PBS medium to remove the Pediococcus pentosaceus cells, subsequently adding to the THP-1 macrophages RPMI 1640 medium with 10% FBS supplemented with 50 μg/ml gentamicin, 10 μg/ml ampicillin and 12 μg/ml chloramphenicol, incubating at 37° C. at 5-7% CO2, and taking aliquots at 5 and 24 hours;
- (h) centrifuging the aliquots and assaying the supernatants for interleukin-10 quantification by flow cytometry;
- and
- (i) calculating the normalized increase of interleukin-10 concentration, with the formula (IL1024h−IL105h)/IL105h;
- wherein IL105h and IL1024h is the concentration of interleukin-10 in μg/ml at 5 and 24 hours, respectively.
Dependencies are inferred from the claim text.